Checking for Oestregen Degradation in Environmental Samples

The aim of this experiment was to identify potential bacteria that utilise oestrogen as a carbon source from any samples obtained. This was achieved through creating a minimal medium with any non-carbon based potential nutrients these bacteria may need, aside from a carbon source – which is provided through 17b-estradiol. The process uses serial dilution over time to do this in order to slowly remove the non-degrading bacteria and allow the degrading bacteria to grow.
1. Add 1ml of sample to 10ml of MDG medium containing 0.1% 17b-estradiol (0.1g) in sterile Erlenmeyer flask (250ml)
2. Add bacteria to shaking incubator and grow at 28°C.
3. Allow bacteria to grow until a dense culture is visible (through cloudiness).
4. Repeat the first method up to 8 times using 100ul of the bacteria that was allowed to grow until a dense culture was visible, do this until there is no reduction in types of colonies formed.
5. After 8 days, form a 101 to 109 serial dilution of samples in physiological saline solution (0.85% NaCl diluted in sterile water).
6. Then add 100ul of each dilution to ISP, R2A and YM culture medium and incubate for 5 days.
7. Any that have grown are capable of oestrogen degradation.

Overall this should identify oestrogen degrading bacteria which can then first be sequenced in order to identified through 16s rRNA sequencing.

This protocol is based on the paper “Degradation of Estrogens by Rhodococcus zopfii and Rhodococcus equi Isolates from Activated Sludge in Wastewater Treatment Plants” by Yoshimoto et al.

@@ Line 5: / Line 5: @@
 		<link rel="stylesheet" href="https://2018.igem.org/Team:Warwick/CSS/GenericPageCSS?action=raw&ctype=text/css">
 		<script src="https://ajax.googleapis.com/ajax/libs/jquery/3.3.1/jquery.min.js"></script>
-<style>.Contents p{font-size: 175% !important}</style>
+<style>.Contents p{font-size: 175% !important} .Contents:nth-child(even){h2: color !important;}</style>
 <script>

Difference between revisions of "Team:Warwick/Experiments"

Revision as of 23:44, 16 October 2018

Experiments

Contact Us

igem@warwick.ac.uk

Sponsors